The domestic honey bee participates in vegetal biodiversity maintenance and food production, since it is one of the most efficient pollinators. These late years, beekeepers happen to face an abnormal level of colony mortality. Neuromuscular studies are needed to better characterize the impact of insecticides on honey bees, since most of active substances on the market are targeting ion channels from insect nerves and muscles, including those acting on the Ryanodine receptor. Electron microscopy and confocal imaging has shown that ultrastructural characteristics of bee skeletal muscle cells resemble those of mammals. Here, excitation-contraction coupling and intracellular calcium signaling was further characterized at the subcellular level in intact skeletal muscle fibers enzymatically isolated from legs of bees aged 1-3 days. Confocal x-y and line-scan observations were taken in a physiological Tyrode’s solution containing calcium, after loading with the calcium indicator fluo-8 AM. Spontaneous localized calcium release events (CRE) were frequently detected. Occasionally, propagating calcium waves were observed. In fibers showing spontaneous activity, event frequency was calculated from x-y sequences taken at 10 Hz and was 2200±470 Hz/mm2 (n=15 cells). Images were automatically processed and gave a preliminary characterization of CRE. They looked ‘wide’ as spatial spread at half maximum was 3.71±0.02 and 3.28±0.02 µm (n=5174 events) parallel with and perpendicular to the fiber axis, respectively. The mean amplitude of the events was 0.220±0.001. Frequency and spatial spread were thus different from events (sparks, embers) detected previously in cardiac myocytes, batrachian and mammalian skeletal muscle fibers. For the first time, subcellular calcium events were monitored in isolated skeletal muscle cells from an arthropod and this new approach may help in understanding their role and regulation in muscles and the myotoxicity of insecticides.